for
Min-Ho Lee
mhlee at WWW.BRIC.POSTECH.AC.KR
Thu Dec 26 01:04:35 EST 1996
Shou Jiang wrote:
>
> Hi, Netters,
>
> Who would like to offer more effective methods to extract lambda DNA from
> lambda library? any kit exist?
>
> Thanks in advance.
>
> Please reply to shouj at nic.bmi.ac.cn
>
> Shou Jiang
Hi!
We collected this subject in our web page
(http://bric.postech.ac.kr/http://bric.postech.ac.kr/forum/bric-forum/newsgroup/bio_methods/)
as follows;
Subject: Re: Lambda DNA extraction protocol
Date: Wed, 04 Dec 1996 | From: robin_beech at maclan.mcgill.ca (Robin
Beech)
slee at pbi.nrc.ca ("Lee, Sun") wrote: Would anyone like to introduce me a
really simple way for this? Most of published methods that were armed
with 'easy' , 'simple' .... were
actually not like that. Looking for in -hand method that really works.
Lambda miniprep
1) Grow an overnight liquid lysate
2) Spin down 1ml of lysate to remove bacterial debris
3) Transfer 0.8ml of lysate to a new eppendorf
4) Add DNAse to 1ug per ml and incubate at RT for 15 min
5) Add 0.2ml of Tris (50mM pH 8.0) EDTA (10mM) SDS (5%) shake
gently and incubate 70 deg for 15 min
6) Cool on ice and add 0.15ml 8M potassium acetate
7) Spin down the protein precipitate
8) Phenol-chloroform extract the supernatent
9) Add 2.5 vol of ethanol or 1 vol of isopropanol to precipitate
the DNA
This can also be scaled up to process 8ml of lysate Hope this helps -
good luck
Robin N. Beech
Institute of Parasitology
McGill University
------------
Min-Ho Lee ; mhlee at bric.postech.ac.kr
Web for the Korean Entomologist (my home page) :
http://bric.postech.ac.kr/about/mhlee/
BRIC, POSTECH
http://bric.postech.ac.kr
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